Allert J. Jonker

Distribution of dopamine in the forebrain and midbrain of the red-eared turtle, Pseudemys scripta elegans, reinvestigated using antibodies against dopamine.

The distribution of dopamine (DA) immunoreactivity in the forebrain and midbrain of the red-eared turtle, Pseudemys scripta elegans, was studied using recently developed antibodies against DA. DA-containing cells were found around the glomeruli of the olfactory bulb but not in the telencephalon proper. In the diencephalon DA cells were observed in the preoptic region, several parts of the periventricular hypothalamic nucleus, the periventricular organ, the ependymal wall of the infundibular recess, the lateral hypothalamic area and the pretectal posterodorsal nucleus. In the midbrain DA cells were found in the ventral tegmental area, the substantia nigra and the presumed reptilian homologue of the mammalian A8 cell group. Dopaminergic fibers and terminals were observed throughout the whole brain, particularly in the telencephalon and diencephalon. The olfactory tubercle, the striatum and the nucleus accumbens appear to have the most dense innervation, but the anterior olfactory nucleus and the septal area also show numerous DA fibers and terminals. Cortical areas are in general not densely innervated by DA fibers. Compared to the results obtained for a lizard, Gekko gecko, with the same antibodies, the results of the present study are very similar as regards the distribution of DA neurons, fibers and terminals. In having better developed DA cell groups in the midbrain and a stronger innervation of the striatum, Pseudemys resembles mammals more than does Gekko. In contrast, the many cerebrospinal fluid-contacting DA neurons in the hypothalamus of Pseudemys are a primitive feature of the diencephalon. The previous immunohistochemical study of Gekko, a lizard, and the present account of Pseudemys, a turtle, indicate that at least two different lines of evolution exist within the reptiles with regard to the DA innervation of the dorsal ventricular ridge. One, including turtles and, probably, crocodilians with a weak DA innervation; and another, represented by lizards, with a strong DA immunoreactivity.

Comparative analysis of vasotocin-like immunoreactivity in the brain of the turtle Pseudemys scripta elegans and the snake Python regius.

The distribution of vasotocin in the brains of the turtle Pseudemys scripta elegans and the snake Python regius was studied with immunohistochemical methods. In both species, vasotocin-immunoreactive (VTi) cells were found in the supraoptic nucleus, the paraventricular nucleus and the bed nucleus of the stria terminalis. No VTi cell bodies were seen in the brainstem. Vasotocinergic fibers were found in all major brain divisions. Intrahypothalamic VTi fibers were observed between the supraoptic and the paraventricular nuclei and in the median eminence. An extensive network of extrahypothalamic VTi fibers extends from the olfactory bulb to the spinal cord. Limbic structures, such as the nucleus accumbens, the septal area and the ventral amygdaloid nucleus, contain a moderate to dense VTi plexus. Other areas with a substantial number of VTi fibers are the lateral habenular nucleus, the ventral tegmental area, the substantia nigra, the locus coeruleus and the nucleus of the solitary tract. Sex-related differences in the density of the VTi fibers were observed in the lateral septal nucleus, the mid-brain periaqueductal gray and, to a lesser extent, in the ventral amygdaloid nucleus, the lateral habenular nucleus, the ventral tegmental area and the substantia nigra. In these areas, the density of VTi fibers is higher in males than in females. The distribution of vasotocin-like immunoreactivity in the brains of Pseudemys and Python resembles the pattern previously observed in the lizard Gekko gecko. However, among the three species several differences exist, the most remarkable one being the variation in number of liquor-contacting VTi cells in the paraventricular nucleus.

Differential effects of dopamine depletion on the binding and mRNA levels of dopamine receptors in the shell and core of the rat nucleus accumbens

In the present study, using quantitative receptor autoradiography and in situ hybridization histochemistry the effects of unilateral 6-hydroxydopamine lesions on the binding density levels of dopamine D1 and D2 receptors and the levels of mRNA encoding D1 and D2 receptors were investigated in the core and shell territories of the nucleus accumbens (Acb) and in the caudate-putamen (CP). The lesions induced contrasting effects on the D1 binding and D1 mRNA in the Acb and CP, i.e. an increase in binding and a decrease in the mRNA levels. For the D2 receptor an increase in both the binding density and mRNA levels was observed. The lesion-induced effects displayed regional differences. For D1 mRNA and D1 and D2 binding, the lesion effect was more pronounced in the core than in the shell of the Acb. For the D2 mRNA levels an increase was observed in the CP but not in the two territories of the Acb. Furthermore, the decrease in D1 mRNA was greater in the rostral than in the caudal parts of the core and shell of the Acb. These results indicate that the core and shell of the Acb and the CP respond differentially to dopamine depletion.

Rostrocaudal Subregional Differences in the Response of Enkephalin, Dynorphin and Substance P Synthesis in Rat Nucleus Accumbens to Dopamine Depletion

Quantitative in situ hybridization histochemistry was used to examine the effects of unilateral 6–hydroxydopamine lesions of the ascending dopaminergic fibres on levels of mRNA encoding the neuropeptides enkephalin, dynorphin and substance P in subregions of the nucleus accumbens. The nucleus accumbens was divided into quadrants and changes in mRNA were measured along the rostrocaudal extent of the nucleus. Two weeks after the lesion an increase was found in enkephalin mRNA in the lesioned side compared to the non‐lesioned side, whereas a decrease was observed for dynorphin and substance P mRNA. The changes in mRNA levels differed from quadrant to quadrant and were not uniformly distributed along the rostrocaudal axis. Both types of changes, i.e. increase and decrease, were much higher in rostral parts of the nucleus than in caudal parts, indicating regional differences in the effects of blockade of the dopaminergic neurotransmission. The lesion‐induced increases and decreases in mRNA levels occurred in both the shell and the core subregions of the nucleus accumbens and were not specifically related to either of these areas. Factors are discussed that may contribute to the rostrocaudal gradient in the changes of enkephalin, substance P and dynorphin mRNA levels. On the basis of their afferent and efferent connections, the rostral and caudal parts of the nucleus accumbens are considered to be involved in different functions. The present results suggest that dopamine depletion may affect these functions in a differential manner.

Compartment-specific changes in striatal neuronal activity during expression of amphetamine sensitization are the result of drug hypersensitivity

Repeated exposure to drugs of abuse induces behavioural sensitization, i.e. a persistent hypersensitivity to the psychomotor stimulant effects of these drugs. This may be the result of increased responsiveness, to drugs, of mesostriatal dopamine systems and their projections, but it has also been suggested that acute and sensitized behavioural responses to psychostimulant drugs involve activation of distinct neuronal circuits. In order to distinguish between these possibilities, we studied amphetamine‐induced c‐fos immunoreactivity in subregions of rat striatum (patch and matrix compartments of caudate–putamen and nucleus accumbens core and shell) in drug‐naive rats, as well as during long‐term expression of amphetamine sensitization. We found that, in sensitized animals, amphetamine (1.0 mg/kg) evoked an increase in the ratio of c‐fos‐immunopositive cells in striatal patch and matrix compartments, suggesting a preferential involvement of striatal patches in the sensitized response to amphetamine. In drug‐naive rats, amphetamine (0.5–5.0 mg/kg) dose‐dependently increased c‐fos expression in all striatal subregions. Remarkably, the highest dose of amphetamine also evoked an increase in patch : matrix ratio of c‐fos immunoreactivity. In nucleus accumbens core and shell of amphetamine‐ and saline‐pretreated animals, amphetamine (1.0 mg/kg) evoked comparable increases in c‐fos expression. These data indicate that distinct striatal compartments display a differential sensitivity to amphetamine in both drug‐naive and amphetamine‐sensitized animals. In addition, they suggest that the shift in amphetamine‐induced c‐fos expression from striatal matrix to patches in sensitized animals is the consequence of a change in the sensitivity to amphetamine, rather than a long‐term circuitry reorganization that is exclusive to the sensitized state.

Morphine-induced increase in D-1 receptor regulated signal transduction in rat striatal neurons and its facilitation by glucocorticoid receptor activation: Possible role in behavioral sensitization

One month (but not 1–3 days) after intermittent morphine administration, the hyperresponsiveness of rats toward the locomotor effects of morphine and amphetamine was associated with an increase in dopamine (DA) D-1 receptor-stimulated adenylyl cyclase activity and enhanced steady state levels of preprodynorphin gene expression in slices of the caudate/putamen and nucleus accumbens. Such an enduring increase in postsynaptic D-1 receptor efficacy also occurred in cultured γ-aminobutyric acid (GABA) neurons of the striatum obtained from rats prenatally treated with morphine. Interestingly, in vitro glucocorticoid receptor activation in these cultured striatal neurons by corticosterone potentiated this neuroadaptive effect of prior in vivo morphine exposure. Since activation of glucocorticoid receptors by corticosterone did not affect D-1 receptor functioning in cultured neurons of saline-pretreated rats, prior intermittent exposure to morphine (somehow) appears to induce a long-lasting state of corticosterone hyperresponsiveness in striatal neurons. Therefore, DA-sensitive striatal GABA neurons may represent common neuronal substrates acted upon by morphine and corticosterone. We hypothesize that the delayed occurrence of these long-lasting morphine-induced neuroadaptive effects in GABA/dynorphin neurons of the striatum is involved in the enduring nature of behavioral sensitization to drugs of abuse and cross-sensitization to stressors.

Differential localization of mRNAs encoding dopamine D1 or D2 receptors in cholinergic neurons in the core and shell of the rat nucleus accumbens

By combining immunocytochemistry for ChAT and in situ hybridization for dopamine D1 or D2 receptor mRNA in the striatum, it was found that (1) the percentage of ChAT/D2 mRNA co-localization is higher in the caudate-putamen than in the shell and core of the nucleus accumbens, (2) in the shell the degree of ChAT/D2 mRNA co-localization is higher rostrally than caudally, and 3) no significant regional differences exist in the degree of co-localization of ChAT and D1 mRNA.

Expression of Enkephalin in Pallido‐Striatal Neurons

Lesions of the ascending dopaminergic system in the rat or neuroleptic treatment may lead to an upregulation of activity in globus pallidus, which is contrary to what would be expected based on current models of basal ganglia function.1 It is not known if such a response occurs in all pallidal neurons or whether subpopulations of pallidal neurons react differently. Pallidal neurons reach several different target nuclei.2 In the present experiments we tried to differentiate between subpopulations of pallidal neurons by determining the presence of prepro-enkephalin mRNA. Next, possible projection targets of the enkephalin-expressing pallidal neurons were identified. Finally, we determined the response of the enkephalin-expressing cells to unilateral dopamine depletion by midbrain 6-hydroxydopamine (6-OHDA) lesion.

C-fos activation patterns in rat prefrontal cortex during acquisition of a cued classical conditioning task. .

The prefrontal cortex (PFC) is known to be involved in associative learning; however, its specific role in acquisition of cued classical conditioning has not yet been determined. Furthermore, the role of regional differences within the PFC in the acquisition of cued conditioning is not well described. These issues were addressed by exposing rats to either one or four sessions of a cued classical conditioning task, and subsequently examining c-fos immunoreactivity in various areas of the PFC. Differences in patterns of c-fos immunopositive nuclei were found when comparing the PFC areas examined. No significant differences were found between rats presented with a temporally contingent conditioned stimulus (CS) light and food (paired groups) and those presented with the same stimuli temporally non-contingently (unpaired groups). In lateral and orbital PFC, both the paired and unpaired groups showed more c-fos immunopositive nuclei than control groups exposed only to the behavioral setup (context exposed groups), and all groups showed a drop in c-fos immunopositive nuclei from session 1 to session 4. In dorsal medial PFC, no differences were seen between the paired, unpaired and context exposed groups. These groups did, however, differ from naive animals, an effect that was not seen in the ventral medial PFC. The results of this study do not support a role for the PFC in the acquisition of a cued classical conditioning task. The differences seen between paired, unpaired and context exposed groups in orbital and lateral PFC could be due to contextual conditioning or reward-related effects.

Alpha-MPT does not affect dopamine levels in the periventricular organ of lizards.

Recent studies indicate that the liquor (CSF)-contacting cell bodies in the periventricular organ of non-mammalian vertebrates accumulates rather than synthesizes dopamine. We therefore gave the lizard Gekko gecko injections of the dopamine synthesis inhibitor alpha-methylparatyrosine (alpha-MPT). Brains were fixed 105-240 min after injection. Subsequent staining with dopamine antiserum revealed almost no changes in staining in the periventricular organ but there was a dramatic decrease in the other dopaminergic cell groups of the brain. The data support the notion that the cells in the periventricular organ accumulate dopamine and that the CSF plays an important role in dopamine neurotransmission in non-mammalian vertebrates.